The quality control method and standard were established to control the quality of Pteris multifida in this paper. The tests of water content, total ash, acid-unsoluble ash and ethanol-soluble extractives of P. multifida were carried out according to the methods recorded in appendix of Chinese Pharmacopeia (2010 edition, volume 1) . The TLC method was established by using rhoifolin as references, and a mixture of CHCl3 -MeOH-HAc (6: 1: 1) as the developing solvent system on GF254 thin layer plate. The contents of rhoifolin was determined by HPLC on a Diamonsil C18 (4.6 mm x 250 mm, 5 μm) column, using acetonitrile-water (containing 0.15% formic acid) (16: 84) as mobile phase at a flow rate of 1.0 mL x min(-1). The column temperature was 30 degrees C and the detection wave-length was 350 nm. As a result, pterosin C 3-O-β-D-glucosidede and the other constituents were well separated on TLC detected under the UV light at 254 nm . The methodology validation for the assay of rhoifolin presented that it was in good linear correlation in the ranges of 0.025 5-5.1 μg with the regression equations of Y = 1 092.4X + 9.503 5 (r = 0.999 8), and the average recoveries were 100.3% (RSD 1.3%). The content range of rhoifolin from 16 different batches of Pteris multifida was 0.08-5.06 mg x g(-1). The water content, total ash, acid-unsoluble ash and ethanol-soluble extractives of 16 samples varied in the ranges of 7.35% - 12.96%, 6.90% - 16.33%, 2.07% -11.38% and 13.29% -23.87%, respectively. The suggesting limes in the quality standard for water content, total ash, acid-unsoluble ash, ethanol-soluble extractives and rhoifolin content were ≤ 12% , ≤ 15% , ≤ 8.5% , ≥ 14% and ≥ 0.040%, respectively. The result proved that the established quality of control method was specific and accurate, which can be used for the quality control of P. multifida.
China ; Chromatography, High Pressure Liquid ; Chromatography, Thin Layer ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; standards ; Pteris ; chemistry ; Quality Control

Objective To construct lentiviral vector miR30-CD133 targeting CD133 gene and investigate its effects on SMMC7721 cells. Methods Thespecific sequence of DNA which containing both sense and antisense Oligo DNA of the targeting sequence were designed, synthesized and cloned into the pPRIME vector. pPRIME-miR30-CD133, psPAX2 and pMD2G were co-transfected on 293T cells to get the lentivirus the transfection efficiency was investigated by confocal laser scanning microscope. The expression of miR30-CD133 on CD133 were detected by qRT-PCR and Western blott. The proliferation and apoptosis effect of miR30-CD133 was respectively evaluated by CCK-8 assay and AnnexinV/PI. Results Functional PFU titers of PRIME-miR30-CD133 were 6.58 × 109 PFU/mL. The expression of CD133 mRNA in sh CD133 group (0.18 ± 0.04) was less than Blank group and shNC group (P<0.05). The expression of CD133 protein in sh CD133 group was less than Blank group (10%) and shNC group (8%) (P < 0.05). Cells proliferation activity were significantly inhibited in shCD133 group compared with control group. Apoptosis rate were significantly increased in shCD133 group (41.3%) compared with Blank group (25.3%)and shNC group (24.3%). Conclusion The lentivirus miR30 vector targeting CD133 gene was successfully constructed, which will be a basis to the further CD133 gene studies.

Objective To isolate and culture the rabbit mesenchymal stem cells,and explore the biological features in vitro.Methods Two milliliter of bone marrow was extracted from the tibia of juvenile rabbit under anesthesia and sterile conditions,and mesenchymal stem cells were obtained by density gradient centrifugation and inoculated to 100ml culture flask.The morphological changes,adherence rate and ultrastructure of the stem cells were observed.The surface markers and growth cycle of the mesenchymal stem cells were detected by flow cytometry,and biological features were observed by inducing the cells differentiate into osteoblasts.Results The adherence of mesenchymal stem cells started 2 hours after culturing,basically finished 20 hours later,and more than 90% of the cells were fused at the 15th day.Observed with transmission electron microscopy,the cultured cells showed typical ultrastructure of stem cells: numbers of microvillus mainly distributed on one side of the cells,irregular nuclei with indentation,and lots of cell organs.Analysis for the cell surface markers indicated positive expression of CD44 and CD90 and negative expression of CD45 in mesenchymal stem cells.Cell growth cycle assay showed that most mesenchymal stem cells(about 97%) stayed in stage G1 and G2.Three weeks after osteoblastic induction,the mesenchymal stem cells gradually produced small calcium salt crystals,the cells were positive in alkaline phosphatase staining,and Von Kossa staining showed many calcium salts in the cell clone as black reaction deposits.Conclusions The rabbit bone marrow mesenchymal stem cells isolated by density gradient centrifugation are of high purity and stable growth rate,and the cells' characters can be still maintained after serial subcultivation.Since easy to operate on isolation and proliferation,the mesenchymal stem cells could be used as the tissue engineering seed cells in repairing of intervertebral disc degeneration.

Objective To apply the oral gastrointestinal ultrasound contrast agents for evaluating the gastric motility abnor‐mality in the patients with anxiety disorder .Methods Twenty patients with anxiety disorder complicating upper digestive tract symptoms without organic pathological changes were enrolled as the anxiety disorder group .Twenty healthy volunteers were en‐rolled as the control group in this study .The two groups orally took gastrointestinal ultrasound contrast agents .The antral contrac‐tion frequency ,antral contraction amplitude amd MI were measured at each time point .GER was calculated .The the gastric motility parameters in the patients with anxiety disorder were evaluated .Results The antral contraction frequency and MI at initial 2 min had no statistical difference between the anxiety group and the control group .The antral contraction amplitude ,antral contraction frequency amd MI at each time point during 5 -10 min after contrast in the anxiety disorder group were significantly decreased compared with the control group ,and the differences were statistically significant (P< 0 .05) .After 20 min ,GER in the control group was significantly higher than that in the anxiety group ,the difference was statistically significant (P<0 .05) .Conclusion O‐ral gastrointestinal ultrasound contrast agents is a economic ,strongly operable ,non‐invasive and highly repeatable method for evalu‐ating the gastric motility .

Objective:To explore the stress change rule of craniofacial bone suture and the interface of bone-implant against differ-ent strength and direction of protraction on the implant anchorage in alveolar bone.Methods:The original DICOMdata of 2-D image of craniofacial complex were obtained by high resolution CT scanning.3-D finite element models of craniofacial complex were devel-oped according to the DICOMdata.Forces of 1 -1 0 N inclined at 0 -60°to Frankfort horizontal plane in the anterior and inferior di-rections were respectively applied on the implant anchorage in the alveolar bone at 32 23 .Data of principle stress and Von Mises Stress of each mode of each simulaton was caculated.Results:The change rule of the effectiveness of different force value of protrac-tion in the same direction was the same in different stress zone;that of the same force value of the protraction in differente direction differed in different stress zone.When the protraction angle was less than 30°,the maxillary complex will spin up.In the 30°,the maxillary complex showed the forward growth.Between 40°-50°,the growth direction was the same with the protraction direction. When the protraction angle was more than 50°,the maxillary complex showed down spin.Conclusion:Protraction force of 1 -1 0 N at 30°-50°to Frankfort horizontal plane on implant anchorage in the alveolar bone at 32 23 can induce maxillary complex grow for-ward.

Background and purpose:With the widespread use of screening of prostate-specific antigen (PSA) levels, prostate cancers at organ-conifned stage are increasing in newly diagnosed cases. However, some defects remain in conventional monoexponential diffusion-weighted imaging (DWI) for differentiating organ-conifned prostate cancer from benign lesions. Therefore, the aim of this study was to obtain biexponential apparent diffusion parameters of prostate organ-conifned cancer, chronic prostatitis in peripheral zone (PZ) and normal PZ tissue, and to compare with monoexponential apparent diffusion coeffcient (ADC) for differentiating prostate cancer from prostatitis lesions. Methods:Sixteen patients with pathologically confirmed prostate organ-confined cancer in PZ, 14 with prostatitis underwent conventional (b-factors 0, 1 000 s/mm2) and 10b-factors (0-3 000 s/mm2) diffusion-weighted imaging (DWI).The monoexponential ADC value and biexponential parameters fast ADC (ADCf), fraction of ADCf (f), slow ADC (ADCs) value for prostate cancer, prostatitis and normal tissues were calculated and compared. Receiver operating characteristic analysis was performed for those parameters.Results:Biexponential and monoexponential parameters were obtained for 18 prostate cancers, 18 prostatitis and 37 normal PZ tissues. The ADC value of prostate cancer tissues was remarkably lower [(0.83±0.11)×10-3 mm2/s] than that of other tissues (P<0.01), while the ADC value of prostatitis [(1.45±0.19)×10-3 mm2/s] was lower than that of PZ [(1.67±0.31)×10-3 mm2/s] (P<0.01). Prostate cancer tissues had low-er ADCf [(1.54±0.23)×10-3 mm2/s],f [(45.8±5.4)%] and ADCs [(0.52±0.15)×10-3mm2/s] than the other tissues (P<0.01). The ADCf,f and ADCs were higher in PZ [(3.90±0.40)×10-3, (67.3±8.2)% and (1.51±0.36)×10-3 mm2/s] than prostatitis [(3.06±0.49)×10-3, (47.9±3.9)% and (0.91±0.29)×10-3 mm2/s) (P<0.01). The area under the curve (AUC) of ADCf and ADC were similar in differentiating cancer and prostatitis (0.96vs 0.94) (P>0.01), but the AUC off and ADCs in differ-entiating cancer from prostatitis (0.83 and 0.80) were signiifcantly lower than that of ADC (P<0.01).Conclusion:The biexponential DWI provided additional tissue characterization parameters for different prostate tissues. ADCf yielded comparable accuracy with ADC in identiifcation of prostate organ-conifned cancer. The biexponential parameter could further improve the diagnostic effcacy.

Objective To investigate the characteristics of memory impairment in patients with ischemic cerebrovascular diseases(ICVD)for further understanding in the cognitive impairment at early stage.Methods Forty-six ICVD patients aged 49 to 80 years old and forty-six matched volunteers,all from the same hospital during Sep 2005 to Feb 2006,were enrolled.Mini-mental state examination(MMSE),clinical memory scale(CMS),Fuld object-memory evaluation(FOM)and rapid verbal retrieve(RVR)were used to evaluate the cognitive and memory function.Results The ICVD patients had lower scores in every memory function than the controls(P

Objective To investigate the influence of age on clinical features and prognosis of severe traumatic brain injury.Methods A total of 135 patients with severe traumatic brain injury were divided into four groups according to age:the juvenile group (＜18 years,15 cases),the young adult group (18 44 years,77 cases),the middle aged group (45 59 years,37 cases) and the elderly group (＞60 years,6 cases).Neurological functions were assessed by the Disability Rating Scale (DRS),the Mini-Mental Status Examination (MMSE),the Fugl-Meyer motor scale (FM) and the Modified Barthel Index (MBI).All the patients were followed up with DRS evaluation 1 4 years after discharge from the hospital.Results MMSE scores decreased with age,with statistically significant differences between the elderly group and the juvenile group,the young adult group and the middle aged group,respectively [(11.0±5.2) vs (21.5±8.1),(21.4±8.0) and (19.1±8.1),respectively; t=2.663、2.825、2.561,P=0.015,0.006,0.022,respectively].Similarly,when compared with the other groups,the elderly group also showed statistically significant differences in follow-up DRS scores [(12.8±6.1) vs.(4.3±3.6),(6.7±5.0) and (7.8±6.9),respectively; t=-2.382、-2.587、-2.385,P =0.040,0.013 and 0.038,respectively]; and the DRS score differentials [(2.3±4.6) vs.(6.2±4.3),(6.7±3.1) and (4.6±3.1); t=2.366、2.242、2.626,P =0.004,0.013 and 0.009,respectively].Conclusions Age may be one of the factors that affect cognitive functions and prognosis associated with traumatic brain injury,and the prognosis for elderly patients is generally unfavorable.

BACKGROUND:Caveolin-1 is expressed in mammalian brain and involved in the normal development of the brain, which can affect the proliferation of neural stem cells in the brain. OBJECTIVE:To acquire neural stem cells from caveolin-1 knockout embryonic mice in vitro and study their biological characteristics. METHODS:The whole brain was separated from C57BL/6 mice and caveolin-1 knockout C57BL/6 mice respectively at encyesis 14-16 days. Single cellsuspension was obtained by enzyme digestion, and cultured in the conditioned medium of neural stem cells. Fol owing 7 days of primary culture, the cells were induced in Dulbecco’s modified Eagle’s medium/Ham’s nutrient mixture F-12 containing 10%fetal bovine serum for 7 days. RESULTS AND CONCLUSION:The major cells of the cellsuspensions from the fetal mouse brain were dead at 1 day after culture, and some single cells floated in the medium and their transmittance were better, and then they gradual y formed multicellular bal s after 3 days. A smal amount of cells were adhered at the bottom of culture plate after passage, and a great amount of cellbal s appeared after 7 days. The proliferation rate of neural stem cells from caveolin-1 knockout mice was higher than that from normal mice. The cellbal s were nestin-positive and their differentiated cells was positive for neurofilament 200, glial fibril ary acidic protein or O4, respectively. Al of the cells from normal mouse brain were positive for caveolin-1, but the cells from caveolin-1 knockout mice were negative for caveolin-1 by immunocytochemistry. Moreover, the speed of cellbal formation and the number of cellbal s in neural stem cells from caveolin-1 knockout mice were better than those from normal mice. Caveolin-1 negative neural stem cells were cultured successful y from caveolin-1 knockout mouse brain, and the results show that caveolin-1 can promote the proliferation of neural stem cells and inhibit their differentiation in vitro.

Acute Disease ; Humans ; Oxalates ; poisoning